An improved assay for platelet-activating factor using HPLC-tandem mass spectrometry. J Lipid Res 2005 Feb;46(2):373-82
Date
11/03/2004Pubmed ID
15520455DOI
10.1194/jlr.D400029-JLR200Scopus ID
2-s2.0-13944253190 (requires institutional sign-in at Scopus site) 20 CitationsAbstract
We describe an improved assay for platelet-activating factor (PAF; 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) using HPLC-tandem mass spectrometry (LC-MS/MS). The present method can readily detect as little as 1 pg (1.9 fmol) of PAF, a significant improvement over previously described LC-MS/MS methods, and gives a linear response up to 1,000 pg of PAF. Our method also overcomes the artifacts from isobaric lipids that have limited the usefulness of certain existing LC-MS/MS assays for PAF. In the course of these studies, we detected three novel lipid species in human neutrophils. One of the novel lipids appears to be a new molecular species of PAF, and the other two have chromatographic and mass spectrometric properties consistent with stearoyl-formyl-glycerophosphocholine and oleoyl-formyl-glycerophosphocholine. These observations identify previously unknown potential interferences in the measurement of PAF by LC-MS/MS. Moreover, our data suggest that the previously described palmitoyl-formyl-glycerophosphocholine is not unique but rather is a member of a new and poorly understood family of formylated lipids.
Author List
Owen JS, Wykle RL, Samuel MP, Thomas MJAuthor
Michael J. Thomas PhD Professor in the Pharmacology and Toxicology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
CatalysisChromatography
Chromatography, High Pressure Liquid
Clinical Chemistry Tests
Glycerophospholipids
Humans
Lipid Metabolism
Lipids
Lysophosphatidylcholines
Mass Spectrometry
Neutrophils
Platelet Activating Factor
Sensitivity and Specificity
Time Factors